教師資料查詢 | 類別: 期刊論文 | 教師: 陳曜鴻 CHEN, YAU-HUNG (瀏覽個人網頁)

標題:Biochemical and structural properties of zebrafish Capsulin produced by Escherichia coli
學年99
學期1
出版(發表)日期2011/01/01
作品名稱Biochemical and structural properties of zebrafish Capsulin produced by Escherichia coli
作品名稱(其他語言)
著者Chou, Chi-yuan; Hsu, Chia-hao; Wang, Yun-hsin; Chang, Min-yen; Chen, Li-chao; Chen, Shu-chun; Chen, Yau-hung
單位淡江大學化學學系
出版者Maryland Heights: Academic Press
著錄名稱、卷期、頁數Protein Expression and Purification 75(1), pp.21-27
摘要Capsulin is one of the transcription factors involved in regulating cell differentiation but its biochemical properties and structural characteristics are still unclear. In the present study, we cloned capsulin from zebrafish, which produces large numbers of transparent embryos and has well-characterized developmental stages. By alignment, the deduced amino acid sequence of zebrafish Capsulin, which contains a putative bHLH motif, shares very high homology to that of other species with an 72–82% identity. Zebrafish Capsulin was also targeted to the nucleus of mammalian cells when overexpressed by transient transfection. In order to characterize the structural and biochemical properties of zebrafish Capsulin, a recombinant zebrafish Capsulin protein was expressed and purified in Escherichia coli. By circular dichroism spectroscopy, Capsulin was shown to be 55% α-helical. The size distribution assay by analytical ultracentrifugation indicated that it existed as a monomer–dimer mixture. The results suggested that the recombinant Capsulin has a well-organized and functional structure. Finally, endogenous Capsulin was distributed mainly in the epicardial cells of zebrafish by immunohistochemistry analysis using antibodies raised against zebrafish Capsulin. The present study not only helps us to comparatively analyze capsulin genes across species, but it also provides valuable structural information for further studies of Capsulin biological function in the future.
關鍵字Analytical ultracentrifugation;Anti-Capsulin antibody;bHLH structure;Circular dichroism;Recombinant Capsulin;Nuclear localization
語言英文
ISSN1046-5928; 1096-0279
期刊性質國外
收錄於SCI
產學合作
通訊作者Chou, Chi-yuan; Chen, Yau-hung
審稿制度
國別美國
公開徵稿
出版型式紙本
相關連結
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